AIMS: Protein S-bacillithiolations are mixed disulfides between protein thiols and the bacillithiol (BSH) redox buffer that occur in response to NaOCl in Bacillus subtilis. We used BSH-specific immunoblots, shotgun liquid chromatography (LC)-tandem mass spectrometry (MS/MS) analysis and redox proteomics to characterize the S-bacillithiolomes of B. subtilis, B. megaterium, B. pumilus, B. amyloliquefaciens, and Staphylococcus carnosus and also measured the BSH/oxidized bacillithiol disulfide (BSSB) redox ratio after NaOCl stress. RESULTS: In total, 54 proteins with characteristic S-bacillithiolation (SSB) sites were identified, including 29 unique proteins and eight proteins conserved in two or more of these bacteria. The methionine synthase MetE is the most abundant S-bacillithiolated protein in Bacillus species after NaOCl exposure. Further, S-bacillithiolated proteins include the translation elongation factor EF-Tu and aminoacyl-tRNA synthetases (ThrS), the DnaK and GrpE chaperones, the two-Cys peroxiredoxin YkuU, the ferredoxin-NADP(+) oxidoreductase YumC, the inorganic pyrophosphatase PpaC, the inosine-5'-monophosphate dehydrogenase GuaB, proteins involved in thiamine biosynthesis (ThiG and ThiM), queuosine biosynthesis (QueF), biosynthesis of aromatic amino acids (AroA and AroE), serine (SerA), branched-chain amino acids (YwaA), and homocysteine (LuxS and MetI). The thioredoxin-like proteins, YphP and YtxJ, are S-bacillithiolated at their active sites, suggesting a function in the de-bacillithiolation process. S-bacillithiolation is accompanied by a two-fold increase in the BSSB level and a decrease in the BSH/BSSB redox ratio in B. subtilis. INNOVATION: Many essential and conserved proteins, including the dominant MetE, were identified in the S-bacillithiolome of different Bacillus species and S. carnosus using shotgun-LC-MS/MS analyses. CONCLUSION: S-bacillithiolation is a widespread redox control mechanism among Firmicutes bacteria that protects conserved metabolic enzymes and essential proteins against overoxidation.
AIMS: Protein S-bacillithiolations are mixed disulfides between protein thiols and the bacillithiol (BSH) redox buffer that occur in response to NaOCl in Bacillus subtilis. We used BSH-specific immunoblots, shotgun liquid chromatography (LC)-tandem mass spectrometry (MS/MS) analysis and redox proteomics to characterize the S-bacillithiolomes of B. subtilis, B. megaterium, B. pumilus, B. amyloliquefaciens, and Staphylococcus carnosus and also measured the BSH/oxidized bacillithiol disulfide (BSSB) redox ratio after NaOCl stress. RESULTS: In total, 54 proteins with characteristic S-bacillithiolation (SSB) sites were identified, including 29 unique proteins and eight proteins conserved in two or more of these bacteria. The methionine synthase MetE is the most abundant S-bacillithiolated protein in Bacillus species after NaOCl exposure. Further, S-bacillithiolated proteins include the translation elongation factor EF-Tu and aminoacyl-tRNA synthetases (ThrS), the DnaK and GrpE chaperones, the two-Cys peroxiredoxin YkuU, the ferredoxin-NADP(+) oxidoreductase YumC, the inorganic pyrophosphatase PpaC, the inosine-5'-monophosphate dehydrogenase GuaB, proteins involved in thiamine biosynthesis (ThiG and ThiM), queuosine biosynthesis (QueF), biosynthesis of aromatic amino acids (AroA and AroE), serine (SerA), branched-chain amino acids (YwaA), and homocysteine (LuxS and MetI). The thioredoxin-like proteins, YphP and YtxJ, are S-bacillithiolated at their active sites, suggesting a function in the de-bacillithiolation process. S-bacillithiolation is accompanied by a two-fold increase in the BSSB level and a decrease in the BSH/BSSB redox ratio in B. subtilis. INNOVATION: Many essential and conserved proteins, including the dominant MetE, were identified in the S-bacillithiolome of different Bacillus species and S. carnosus using shotgun-LC-MS/MS analyses. CONCLUSION: S-bacillithiolation is a widespread redox control mechanism among Firmicutes bacteria that protects conserved metabolic enzymes and essential proteins against overoxidation.
Authors: Jason Gioia; Shailaja Yerrapragada; Xiang Qin; Huaiyang Jiang; Okezie C Igboeli; Donna Muzny; Shannon Dugan-Rocha; Yan Ding; Alicia Hawes; Wen Liu; Lesette Perez; Christie Kovar; Huyen Dinh; Sandra Lee; Lynne Nazareth; Peter Blyth; Michael Holder; Christian Buhay; Madhan R Tirumalai; Yamei Liu; Indrani Dasgupta; Lina Bokhetache; Masaya Fujita; Fathi Karouia; Prahathees Eswara Moorthy; Johnathan Siefert; Akif Uzman; Prince Buzumbo; Avani Verma; Hiba Zwiya; Brian D McWilliams; Adeloa Olowu; Kenneth D Clinkenbeard; David Newcombe; Lisa Golebiewski; Joseph F Petrosino; Wayne L Nicholson; George E Fox; Kasthuri Venkateswaran; Sarah K Highlander; George M Weinstock Journal: PLoS One Date: 2007-09-26 Impact factor: 3.240
Authors: Irina V Mikheyeva; Jason M Thomas; Stacey L Kolar; Anna-Rita Corvaglia; Nadia Gaϊa; Stefano Leo; Patrice Francois; George Y Liu; Mamta Rawat; Ambrose L Cheung Journal: Mol Microbiol Date: 2019-02-17 Impact factor: 3.501
Authors: Jordan G Sheppard; Jeremy P McAleer; Pushkar Saralkar; Werner J Geldenhuys; Timothy E Long Journal: Eur J Med Chem Date: 2017-10-12 Impact factor: 6.514
Authors: Vu Van Loi; Manuela Harms; Marret Müller; Nguyen Thi Thu Huyen; Chris J Hamilton; Falko Hochgräfe; Jan Pané-Farré; Haike Antelmann Journal: Antioxid Redox Signal Date: 2016-08-11 Impact factor: 8.401